Luteolin inhibits spike protein of severe acute respiratory syndrome coronavirus‐2 (SARS‐CoV‐2) binding to angiotensin‐converting enzyme 2

Abstract

AbstractSevere acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) causes coronavirus disease 2019 (COVID‐19), a respiratory illness that poses a serious threat to global public health. In an essential step during infection, SARS‐CoV‐2 uses the receptor‐binding domain (RBD) of the spike (S) protein to engage with angiotensin‐converting enzyme 2 (ACE2) in host cells. Chinese herbal medicines and their active components exhibit antiviral activity, with luteolin being a flavonoid that can significantly inhibit SARS‐CoV infection. However, whether it can block the interaction between the S‐protein RBD of SARS‐CoV‐2 and ACE2 has not yet been elucidated. Here, we investigated the effects of luteolin on the binding of the S‐protein RBD to ACE2. By employing a competitive binding assay in vitro, we found that luteolin significantly blocked the binding of S‐protein RBD to ACE2 with IC50 values of 0.61 mM, which was confirmed by the neutralized infection with SARS‐CoV‐2 pseudovirus in vivo. A surface plasmon resonance‐based competition assay revealed that luteolin significantly affects the binding of the S‐protein RBD to the ACE2 receptor. Molecular docking was performed to predict the binding sites of luteolin to the S‐protein RBD‐ACE2 complex. The active binding sites were defined based on published literature, and we found that luteolin might interfere with the mixture at residues including LYS353, ASP30, and TYR83 in the cellular ACE2 receptor and GLY496, GLN498, TYR505, LEU455, GLN493, and GLU484 in the S‐protein RBD. These residues may together form attractive charges and destroy the stable interaction of S‐protein RBD‐ACE2. Luteolin also inhibits SARS‐CoV‐2 spike protein‐induced platelet spreading, thereby inhibiting the binding of the spike protein to ACE2. Our results are the first to provide evidence that luteolin is an anti‐SARS‐CoV‐2 agent associated with interference between viral S‐protein RBD‐ACE2 interactions.